Resistance of B16 melanoma cells to CD47-induced negative regulation of motility as a result of aberrant N-glycosylation of SHPS-1

J Biol Chem. 2004 Apr 2;279(14):13711-20. doi: 10.1074/jbc.M310276200. Epub 2004 Jan 21.

Abstract

The adhesion receptor SHPS-1 activates the protein-tyrosine-phosphatase SHP-2 and thereby promotes integrin-mediated reorganization of the cytoskeleton. SHPS-1 also contributes to cell-cell communication through association with CD47. Although functional alteration of SHPS-1 is implicated in cellular transformation, the role of the CD47-SHPS-1 interaction in carcinogenesis has been unclear. A soluble SHPS-1 ligand (CD47-Fc) has now been shown to bind to Melan-a non-tumorigenic melanocytes but not to syngeneic B16F10 melanoma cells. Treatment of B16F10 cells with 1-deoxymannojirimycin, which prevents N-glycan processing, restored the ability of SHPS-1 derived from these cells to bind CD47-Fc in vitro, indicating that aberrant N-glycosylation of SHPS-1 impairs CD47 binding in B16F10 cells. CD47-Fc inhibited the migration of Melan-a cells but not that of B16F10 cells. However, a monoclonal antibody that reacts with SHPS-1 on both Melan-a and B16F10 cells inhibited the migration of both cell types similarly. CD47 binding induced proteasome-mediated degradation of SHPS-1 in a tyrosine phosphorylation-independent manner. Furthermore, overexpression of SHPS-1 reduced the level of tyrosine phosphorylation of focal adhesion kinase, and this effect was reversed by CD47 binding. These results suggest that CD47 binds to and thereby down-regulates SHPS-1 on adjacent cells, resulting in inhibition of cell motility. Resistance to this inhibitory mechanism may contribute to the highly metastatic potential of B16 melanoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / genetics
  • Antigens, CD / metabolism*
  • Antigens, Differentiation*
  • CD47 Antigen
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Communication / physiology
  • Cell Line, Tumor
  • Cell Movement / physiology*
  • Cysteine Endopeptidases / metabolism
  • Down-Regulation / physiology
  • Focal Adhesion Kinase 1
  • Focal Adhesion Protein-Tyrosine Kinases
  • Glycosylation
  • Ligands
  • Macrophages / cytology
  • Macrophages / metabolism
  • Melanocytes / cytology
  • Melanocytes / metabolism
  • Melanoma, Experimental*
  • Membrane Glycoproteins / metabolism*
  • Mice
  • Multienzyme Complexes / metabolism
  • Neural Cell Adhesion Molecule L1 / metabolism*
  • Phosphorylation
  • Proteasome Endopeptidase Complex
  • Protein-Tyrosine Kinases / metabolism
  • Receptors, Immunologic / metabolism*
  • Skin Neoplasms*
  • Solubility
  • Tyrosine / metabolism

Substances

  • Antigens, CD
  • Antigens, Differentiation
  • CD47 Antigen
  • Carrier Proteins
  • Cd47 protein, mouse
  • Ligands
  • Membrane Glycoproteins
  • Multienzyme Complexes
  • Neural Cell Adhesion Molecule L1
  • Ptpns1 protein, mouse
  • Receptors, Immunologic
  • Tyrosine
  • Protein-Tyrosine Kinases
  • Focal Adhesion Kinase 1
  • Focal Adhesion Protein-Tyrosine Kinases
  • Ptk2 protein, mouse
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex